Agent for reducing the useable calorie content of food and for therapeutic reduction of weight, in particular for use in the case of adiposity (obesity)

ABSTRACT

There are disclosed compositions and methods for treating obesity using 5-D-fructose dehydrogenase. Other embodiments are also described.

This application is a continuation of U.S. Ser. No. 12/094,648, which isa 35 U.S.C. §371 application of PCT/EP2006/011231, filed Nov. 23, 2006,and claims the benefit under 35 U.S.C. §120 of said PCT application, andfurther claims the benefit under 35 U.S.C. §119(e) of U.S. ProvisionalPatent Applications U.S. Ser. No. 60/757,413, filed on Jan. 10, 2006 andU.S. Ser. No. 60/831,173, filed on Jul. 17, 2006. The contents of theseapplications are incorporated herein by reference.

The present invention relates to an agent for reducing the useablecalorie content of food. The agent contains a compound that can effectthe conversion of fructose to 5-keto-D-fructose. 5-keto-D-fructosecannot be metabolized by the human or animal body and is thereforesignificantly less caloric than fructose. The present invention alsorelates to an agent which, in addition to the first compound, contains acompound that can effect the conversion of glucose to fructose.According the invention the term “agent” includes a pharmaceuticalcomposition, a medical device, a foodstuff and a special foodstuff.

According to the present invention, the terms “food” and “foodstuff” areused as synonyms. They mean to also include feed in the sense of animalfeed. In the context of this application “special foodstuffs” arefoodstuffs for particular nutritional uses, foods for special medicalpurposes, medical foods, food supplements, dietary supplements, dieteticfood supplements, health foods, nutraceuticals, and food additives. Inthe context of this application the term foodstuff means to includespecial foodstuffs as used herein, where applicable.

Fructose is a ketohexose and is an important energy providing ingredientof food. It is present as a component of many di- and oligosaccharides,as free fructose or as both in numerous foodstuffs. In contrast toglucose, fructose is assimilated into the mucosa cells of the smallintestine by eased carrier-mediated diffusion. The enzymatic degradationstarts in the liver by the action of the adenosine triphosphate (ATP)dependent fructokinase, whereby fructose is converted to fructose1-phosphate. In the liver and in the kidneys, fructose 1-phosphate iscleaved to glycerine aldehyde and dihydroxy acetone phosphate byaldolase B.

Food, e.g. fruits and fruit juices, contain a large amount of fructose,but in particular also sucrose, which is cleaved to fructose and glucosein the body. Over the past decades, there has been a dramatic increasein the consumption of free fructose (which is approx. 1.6-fold sweeterthan glucose or sucrose) since the cheaper sweetener “high fructose cornsyrup” (HFCS) was added to many of our beverages, bakery products andother sweet foodstuffs. Data from the USA show a parallel developmentbetween the sharp increase of obesity and the additives of freefructose. In contrast to glucose, fructose is metabolized independentlyof insulin. Since insulin influences the occurrence of the sensation ofsatiation indirectly, i.e. fructose does not eliminate the appetite,obesity may easily occur as a result of the extensive use of HFCS as asweetener. Free fructose in large amounts may also favor hypertension.Large amounts of free fructose also influence the lipid profile (bloodlipids) in an unfavorable way, since they promote the synthesis oflipids and thus increase the postprandial serum triglycerides. Patientswith metabolic syndrome should not consume beverages that are sweetenedwith HFCS or sucrose. Fructose is also discussed as a cause of metabolicsyndrome.

Thus, fructose is an important source of calories for the human body andmust be given special consideration in all efforts to reduce the contentof calories in the food. In the following, the term “fructosecontaining” refers to all substances and foodstuffs that either containfructose in pure form or from which fructose can be released in thedigestive tract. The fructose content of substances and foodstuffsrefers to all the fructose in a fructose containing food or substance inwhatever form (e.g. also as part of sucrose) it is contained in such afood or substance.

Departures from normal weight due to an increase in body weight,primarily in the proportion of fat, are referred to as adiposity(obesity), whereby, normally and in the sense of this invention, fourgrades are differentiated, namely grade 0=normal weight, gradeI=overweight, grade II=adiposity and grade III=extreme adiposity. Theindividual grades are classified according to the well-known body massindex (BMI). An increase in fatty tissue and thus in body weight above acertain limit leads to an increase in various diseases, such ashypertension, and life expectancy decreases. The number of personssuffering from adiposity grade I to grade III has increased constantlyin the industrialized countries over the past decades and amounts to 30to 50% of the total population at present, depending on the statisticused and definition of the term overweight or obesity. For example, inthe Federal Republic of Germany, every second person is overweight(adiposity grade I) and every sixth person has adiposity grade II orIII. The number of adipose children is also increasing as a result ofpoor nutrition and lack of exercise. To date, therapy has been limitedto nutrition therapy and, in extreme cases, surgical measures. It is awell-known fact that there is a very large and constantly increasingnumber of nutrition therapies and diets that generally do not result inlasting success. Even if there is an initial loss in weight, usuallythis results in the so-called yoyo effect, i.e. the sufferers initiallylose weight, but put it back on again after the end or early terminationof the diet. Often, in the end the body weight is even higher than priorto the beginning of the diet. Surgical measures, which are only used inparticularly severe cases, have the aim of reducing the supply ofnutrients or nutrient utilization, and are associated with highoperation risks and long-term negative side effects. Nutrient deficitsmay thus result, due to impairments of absorption and insufficientsupply with the food. These relate to vitamins, minerals and proteins.Up to one third of the surgical patients show a folic acid deficiency.In addition, iron, potassium, magnesium and vitamin A deficiencies mayoccur.

The drug therapies known to date for treating adiposity arecontroversial. Products have often had to be taken off the market due tosevere side effects. All drugs that are still available today also havesignificant, unpleasant and frequent side effects, such as fatty stools,constipation and insomnia. A diet that reliably leads to a sustainablesuccess and/or a broadly applicable therapy for adiposity in itsdifferent types without side effects is not known. Of course, thewidespread use of surgical measures does not come into question. Itremains to be said that those suffering from obesity and the specialiststreating them are confronted by a quite hopeless and frustratingsituation for all concerned.

An agent that does not have an effect on the body, but on thecarbohydrates in the food and that would reduce the useable caloriecontent in the food would satisfy an extremely widespread and pressingneed which has existed for decades. Such an agent would also overcomethe prejudice widely held in the specialist world and among thosesuffering from obesity that a lasting weight reduction or the preventionof an increase in weight is not possible without suffering and without asignificant change of nutrition habits. It would mean a dramaticimprovement in the options available for the treatment of adiposity.Such an agent would also put an end to the as yet fruitless efforts ofthe specialist world to find an agent to treat adiposity which can beadministered broadly, easily, and long-term, without causing sideeffects. This would apply all the more to an agent which, in addition,has no negative effects on health.

Thus, there is provided in accordance with embodiments of the presentinvention an agent that significantly reduces the useable caloriecontent of food, in particular for facilitating the intake of foodstuffsthat normally contain fructose also in the case of adiposity, withoutresulting in an increase in weight. Further, there is provided inaccordance with embodiments of the invention an agent to make itpossible for persons suffering from adiposity to eat foodstuffs whichuntil now were not allowed to them due to their fructose content or theeating of which was associated with negative effects for the sufferers'health. There is also provided in accordance with embodiments of theinvention an agent that, after the intake of fructose, can reduce orprevent the associated negative effects on weight and/or health, inparticular in the case of adiposity.

There is also provided, in accordance with embodiments of the inventionan agent that can prevent persons who up to now have not suffered fromadiposity from putting on weight as a result of eating fructosecontaining substances or foodstuffs. There is thus also provided inaccordance with embodiments of the invention an agent to allow personswho are not suffering from adiposity to eat foodstuffs the intake ofwhich would otherwise lead to the risk of weight gain, with theconsequence that these persons might become adipose.

Therefore, the subject matter of the invention is an agent that contains5-D-fructose dehydrogenase (syn. fructose 5-dehydrogenase). The agentaccording to the present invention brings about the conversion offructose in the food into 5-keto-D-fructose by dehydrogenation. Thus,the fructose is changed in such a manner that it is no longer availableto the metabolism of the human or animal body.

According to another aspect of the invention 5-D-fructose dehydrogenaseis used, optionally in combination with glucose isomerase, for treatmentof adiposity, for example in the form of a pharmaceutical composition.The treatment of adiposity may be therapeutic or prophylactic. Also,further diseases accompanying adiposity may be treated by making use ofthe agent according to the invention.

According to another aspect of the present invention, the agent is usedfor lowering the usable calorie content of food.

A subject matter of the invention is also an agent that reduces thebioavailability of fructose in the human or animal body with the help ofa 5-D-fructose dehydrogenase.

A subject matter of the invention is also an agent that reduces thebioavailability of fructose and glucose in the human or animal body withthe help of a 5-D-fructose dehydrogenase in combination with a glucoseisomerase.

A subject matter of the invention is also an agent that reduces thecontent of fructose in a foodstuff with the help of a 5-D-fructosedehydrogenase.

A subject matter of the invention is also an agent that reduces thecontent of fructose and glucose in a foodstuff with the help of a5-D-fructose dehydrogenase in combination with a glucose isomerase.

A subject matter of the present invention is further an agent forreducing the useable calorie content of food which contains a5-D-fructose dehydrogenase—alone or in combination with a glucoseisomerase.

The agent according to the invention is suited for the use in the caseof adiposity and for the therapeutic or prophylactic treatment ofadiposity, possibly accompanied by a further disease usually appearingtogether with adiposity, including diseases of the cardiovascular system(e.g. hypertension, coronary heart disease, venous insufficiency, heartfailure, left-ventricular hypertrophy, arteriosclerosis), of metabolicand hormonal function (e.g. diabetes mellitus type II, dyslipidemias,hyperuricemia, hyperlipoproteinemia), of the respiratory system (e.g.sleep apnea, Pickwickian syndrome), of the hepatobiliary system (e.g.fatty liver, cholecystolithiasis), of the locomotor system (e.g.gonarthrosis, heel spur, arthrosis of the ankle joint), of the skin(e.g. intertrigo, hirsutism, striae) and for use in the case ofneoplasias associated with adiposity (e.g. increased risk ofendometrial, breast, cervical, gall bladder cancer, etc.), disorders ofsexual function (e.g. reduced fertility, complications during birth),psychosocial problems (e.g. reduced self-confidence, social isolation,discrimination, problems with one's partner or at work), and otherproblems, such as reduced mobility and staying power, increased riskduring operations and more difficult examination conditions.

All these diseases and health problems are often associated withadiposity.

A further subject matter of the invention is the use of a 5-D-fructosedehydrogenase—alone or in combination with a glucose isomerase—in thecase of adiposity or in the case of health problems or diseasesassociated with adiposity (see above).

A further subject matter of the invention is the use of 5-D-fructosedehydrogenase—alone or in combination with a further enzyme, preferablytogether with glucose isomerase—for the manufacture of a medicament(pharmaceutical composition) for the prophylactic or therapeutictreatment of adiposity or diseases associated with adiposity (seeabove).

According to the present invention, a 5-D-fructose dehydrogenase—aloneor in combination with a glucose isomerase—may also be used for reducingthe useable calorie content in a foodstuff.

A 5-D-fructose dehydrogenase, in the context of this application, is anenzyme that can catalyze the dehydrogenation of fructose to5-keto-D-fructose.

A possible method for the production of a 5-D-fructose dehydrogenase is,for example, described in Ameyama et al., Journal of Bacteriology 1981,814-823, “D-Fructose Dehydrogenase of Gluconobacter industrius:Purification, Characterization and Application of EnzymaticMicrodetermination of D-Fructose”, the content of which is incorporatedherein by reference.

The practical meaning of such an agent for the reduction of the useablecalorie content of food becomes clearer if one considers that fructoseis used very widely in the foodstuff industry as a sweetener. In thecase of foodstuffs which contain sucrose, the content of calories can bereduced by about one half by the use of the agent according to thepresent invention which contains the enzyme 5-D-fructose dehydrogenase.During digestion, sucrose is enzymatically cleaved to glucose andfructose. The fructose fraction (50% of the calories contained insucrose) is then dehydrogenated by said 5-D-fructose dehydrogenase to5-keto-D-fructose and can therefore no longer be utilized by the body.

Glucose is the central carbohydrate of the energy and substratemetabolism. It is the basic component of many polysaccharides (e.g.starch). In the context of this application the term “glucosecontaining” refers to all substances and foodstuffs that either containglucose in pure form or from which glucose can be released in thedigestive tract. The glucose content of substances and foodstuffs refersto all the glucose in a glucose containing food or substance in whateverform (e.g. also as part of sucrose) it is contained in such a food orsubstance.

Therefore, a further reduction of the calorie content of food may beachieved according to the present invention by an agent that containsglucose isomerase as an additional active ingredient. The invention thusprovides compositions and methods that can be used to enable sufferersof Adiposity to ingest fructose-containing foods or fructose- andglucose-containing foods

A glucose isomerase in the context of this application is an enzyme thatis able to transform glucose into fructose. This conversion can also bebrought about, for example, by a xylose isomerase. Thus, such a xyloseisomerase is, in the context of this application, also a glucoseisomerase. A possible method for the production of a xylose isomeraseis, for example, described in Yamanaka, Biochimica et Biophysika Acta,Volume 151 (3), 1968, 670-680, “Purification, Crystallization andProperties of the D-Xylose Isomerase from Lactobacillus brevis” and inYamanaka, Methods in Enzymology, Volume 41, 1971, 466-471, “D-XyloseIsomerase from Lactobacillus brevis”, the contents of which areincorporated herein by reference.

Such a combination agent can also be used in the form of two separatedose units, e.g. in two separate tablets, one of which contains glucoseisomerase and the other 5-D-fructose dehydrogenase.

By combining 5-D-fructose dehydrogenase with the enzyme glucoseisomerase, ingested glucose as well as the large amounts of glucosereleased by the degradation of carbohydrates are largely removed fromthe metabolism of calories by the body. Glucose isomerase has theproperty of converting glucose into fructose and vice versa with anequilibrium concentration of approximately 50% glucose and 50% fructose.Such an enzyme combination consisting of 5-D-fructose dehydrogenase andglucose isomerase is ideally suited for considerably reducing thecalorie content of, for example starch containing food, such aspotatoes. The glucose that is released from starch during digestion istransformed into fructose by said glucose isomerase, which is thenconverted by said 5-D-fructose dehydrogenase into 5-keto-D-fructosewhich is significantly less bioavailable. Thus, the 5-D-fructosedehydrogenase prevents the above described equilibrium from beingestablished. Therefore, glucose isomerase will convert glucose intofructose, which itself will be dehydrogenated by the 5-D-fructosedehydrogenase to 5-keto-D-fructose, until no further glucose is presentin the food and food pulp.

Also in the case of sucrose, an increased reduction of calories can beachieved with the combination agent. Fructose, which is released fromsucrose during digestion, is converted into 5-keto-D-fructose by5-D-fructose dehydrogenase, as described above. Therefore the glucoseisomerase will attempt to re-establish the above described equilibriumby converting glucose into fructose. This conversion process continuesuntil no further glucose is present in the food pulp. In this way, thecontent of calories of the sucrose containing dishes available to thebody is reduced to a greater degree than if 5-D-fructose dehydrogenaseis used on its own.

The combination agent according to the present invention thus leads to alarge proportion of the glucose supplied to the body or released in thedigestive tract being converted into fructose which in turn is convertedinto 5-keto-D-fructose, which is of much less value for the metabolism.

In a particularly easy way, the invention facilitates the transformationof fructose in a foodstuff into a form that prevents weight gain andenables weight loss. Thus, the invention also facilitates the intake offoodstuffs by persons who suffer from adiposity that had to be avoidedby the sufferers up to now because of their content of fructose andglucose. Further, the invention allows the intake of fructose andglucose containing foodstuffs by persons who are not suffering fromadiposity, without this leading to negative effects on their weightand/or health.

According to the present invention, 5-D-fructose dehydrogenase—alone orin combination with glucose isomerase—is further mentioned for use inmedicine (first medical indication), for example as a pharmaceuticalcomposition. Accordingly, a subject matter of the invention is also aproduct which consists of 5-D-fructose dehydrogenase—alone or incombination with glucose isomerase—or contains 5-D-fructosedehydrogenase—alone or in combination with glucose isomerase—beside oneor more other active ingredients for use in a medical method, inparticular in a method for the therapeutic treatment of the human oranimal body. In the context of this application, a pharmaceuticalcomposition is a product, in particular a substance or a substancemixture, for use in a method for surgical or therapeutic treatment ofthe human or animal body and in diagnostic methods that are performed onthe human or animal body. Thus, in the context of this application,pharmaceutical compositions are also products, in particular substancesor substance mixtures, that are meant or suitable for curing,alleviating, preventing or determining adiposity.

The term “treating” when used in connection with the foregoing disordersincludes amelioration, prevention or relief from the symptoms and/oreffects associated with these disorders and includes the prophylacticadministration of an enzyme or a mixture thereof to diminish thelikelihood or seriousness of the conditions.

According to a further aspect of the present invention, a foodstuff isprovided which contains 5-D-fructose dehydrogenase—alone or incombination—with glucose isomerase. Further, according to the presentinvention, a foodstuff is provided which contains 5-D-fructosedehydrogenase—alone or in combination with glucose isomerase—in anamount which is sufficient to convert fructose into 5-keto-D-fructoseand which, in addition, contains glucose isomerase in an amount which iseffective for transforming glucose into fructose. Such a foodstuff maybe produced using a method for treating a foodstuff in which thefoodstuff is placed in contact with a 5-D-fructose dehydrogenase—aloneor in combination with glucose isomerase—under such conditions underwhich the 5-D-fructose dehydrogenase can dehydrogenate fructose to5-keto-D-fructose and under which the glucose isomerase can convertglucose into fructose. In contrast to otherwise untreated foodstuffs,such a foodstuff has a reduced content of fructose or, if the agentcontains both enzymes, a reduced content of fructose and glucose, andtherefore, for the first time, is suitable to be consumed by persons whowant to control their weight, including adipose persons. Furthermore, afoodstuff can be prepared by a method in which a 5-D-fructosedehydrogenase is added to the foodstuff—alone or in combination withglucose isomerase—in a manner in which the action of the enzyme or bothenzymes only starts after the intake of the foodstuff. Such a foodstuffthat contains 5-D-fructose dehydrogenase or 5-D-fructose dehydrogenaseand glucose isomerase has the same taste as an untreated foodstuff andis, for the first time, suitable to be consumed by persons who want tocontrol their weight including adipose persons due to the reducedcontent of fructose or fructose and glucose which is established aftereating.

According to a further aspect, according to the present invention,5-D-fructose dehydrogenase—alone or in combination with glucoseisomerase—is provided as a medical device. The subject matter of theinvention is accordingly also a medical device that consists of5-D-fructose dehydrogenase—alone or in combination with glucoseisomerase—or contains the 5-D-fructose dehydrogenase—alone or incombination with glucose isomerase—beside one or more other activeingredients.

In the following, the invention will be described further in its variousaspects.

5-D-Fructose dehydrogenase is a compound that has been known for nearly40 years, but has only been used for analytical purposes to date.Glucose isomerase is a compound that has been known for more than 40years and has only been used for starch saccharification to date. In theindustry, it is used for the conversion of glucose into fructose as wellas for the conversion of fructose into glucose.

5-D-fructose dehydrogenase, in particular in combination with glucoseisomerase, has not been used in the medical/pharmaceutical field todate, in particular not in adiposity in humans or animals. Thus, theinvention discloses the first medical indication for 5-D-fructosedehydrogenase and the first medical indication for the combination ofthe two enzymes. Until now, the two enzymes have not been used for thetherapeutic treatment of the human or animal body or for diagnosticpurposes on the human or animal body.

The agent according to the present invention may be taken orally priorto meals, immediately before meals, with meals or immediately aftermeals, so that it can exert its dehydrogenating effect on fructose andoptionally converting effect on glucose in the food pulp. The agentaccording to the present invention may contain the enzyme(s) withoutfurther additives. However, it is preferable that the agent according tothe present invention further contains additives that arepharmaceutically acceptable and/or acceptable for foodstuffs, such asfor example extenders, binders, stabilizers, preservatives, flavourings,etc. Such additives are commonly used and well known for the productionof pharmaceutical compositions, medical devices, foodstuffs, and specialfoodstuffs and the person skilled in the art knows which additives inwhich amounts are suitable for certain presentation forms. The agentsaccording to the present invention may for example contain as additivesdicalcium phosphate, lactose, modified starch, microcrystallinecellulose, maltodextrin and/or fibersol.

The agents according to the present invention can also be added to afoodstuff before eating. They may even be added to the foodstuff at theproduction stage, with the aim of developing their effect only afterconsuming the foodstuff. This may be achieved by microencapsulation, forexample. With this, the useable fructose and/or glucose content of thefoodstuff may be reduced, without negatively affecting its taste.Therefore, preparations are particularly useful that contain5-D-fructose dehydrogenase—alone or in combination with glucoseisomerase—and which release these enzymes only in the digestive tract ofa human or animal or let them become effective in another way, inparticular in the stomach or small intestine. Therefore, the inventioncan be used for example in the production of sweets, fruit preparations(e.g. apple sauce), jam, honey, chocolate and chocolate products, bakeryproducts (e.g. biscuits and cakes), breads, pastas, vegetable dishes,potato dishes, ice cream, cereals, dairy products (e.g. fruit yogurt andpudding), fructose-and/or glucose-containing beverages, fructose- and/orglucose-containing sauces (e.g. tomato ketchup) and fructose- and/orglucose-containing sweeteners. For dishes that are boiled or baked, theagents according to the present invention could e.g. be mixed into orsprinkled onto them after cooling.

Since fructose is widely used as a sweetener in foodstuffs that areespecially produced for diabetics, the addition of the agents accordingto the present invention to diabetic food before eating or the additionof the agents according to the present invention during the productionof diabetic food allows diabetics who suffer from adiposity to eatdiabetic food, such as the above mentioned foodstuffs in theirrespective form as diabetic foods.

The agent according to the present invention may also be added to afoodstuff, to exert its effect on the fructose originating from anotherfoodstuff and in the case of an agent containing both enzymes also onthe glucose originating from another foodstuff after their consumption.An example of this would be the addition of the agent according to thepresent invention to a spread so that the reduction of the calories thatare contained in the bread and that can be utilized by the body occursafter the intake of the bread, without an impairment of taste of thebread. Another example would be mixed spices and mayonnaise, among otherthings for use with french fries.

5-D-fructose dehydrogenase and the combination agent may also be used inimmobilized form. This is useful for the treatment of liquid foodstuffs.For example, 5-D-fructose dehydrogenase can be embedded in a matrixwhich is permeable for fructose. If a fructose containing liquidfoodstuff is allowed to flow along the enzyme containing matrix, thenfructose is extracted from the foodstuff by the action of the enzyme andconverted into 5-keto-D-fructose.

A subject matter of the present invention are also agents that inaddition to other active ingredients also contain 5-D-fructosedehydrogenase, either alone or in combination with glucose isomerase.

The agent may be formulated in any form which is suitable for theintended route of administration. A preferred route of administration isoral administration. For oral administration, the agent may beformulated for example in the form of capsules (coated or noncoated)containing powder, coated or non-coated pellets, granules ormicro-/mini-tablets or in the form of tablets (coated or non-coated)pressed from powder, coated or non-coated pellets, dragées ormicro-/mini-tablets. The agent may also be formulated for example in theform of gel caps or in liquid form as solution, drops, suspension orgel. The agent may also be formulated e.g. as dried or moist oralsupplement. The formulation of the agent according to the presentinvention as powder is particularly suitable for admixing withfoodstuff. The powder may be sprinkled onto a meal or mixed into a pulpor beverage. It is particularly beneficial, if the agent offered as bulkpowder is packaged in single dosage amounts, such as in single bags orcapsules, or if it is provided in a dosing dispenser.

For oral administration, the 5-D-fructose dehydrogenase—alone or incombination with glucose isomerase—may be used with acceptableexcipients and/or carriers.

The total amount of the carrier and/or excipient of an agent containing5-D-fructose dehydrogenase or 5-D-fructose dehydrogenase and glucoseisomerase is preferably between 5 and 99.9% by weight, more preferablybetween 10 and 80% by weight and even more preferably between 25 and 60%by weight of the composition.

Suitable excipients and/or carriers include maltodextrin, calciumcarbonate, dicalcium phosphate, tricalcium phosphate, microcrystallinecellulose, dextrose, rice flour, magnesium stearate, stearic acid,croscarmellose sodium, sodium starch glycolate, crospovidone, sucrose,vegetable gums, lactose, methylcellulose, povidone, carboxymethylcellulose, corn starch, modified starch, fibersol, gelatine,hydroxypropylmethyl cellulose and the like (including mixtures thereof).

Preferable carriers include calcium carbonate, magnesium stearate,maltodextrin, dicalcium phosphate, modified starch, microcrystallinecellulose, fibersol, gelatine, hydroxypropylmethyl cellulose andmixtures thereof.

The various ingredients and the excipient and/or carrier may be mixedand formed into the desired form using common methods well known to theskilled person. The administration form according to the presentinvention which is suited for the oral route, such as e.g. tablet orcapsule, may be coated with a coating which is resistant against low pHvalues (approximately pH 1 to 2.5) and which dissolves at a pH value ofapproximately 3.0 to 8.0, preferably at a pH value of 3.0 to 6.5 andparticularly preferable at a pH value of 4.0 to 6.0. An optionally usedcoating should be in accordance with the pH optimum of the enzyme usedand its stability at pH values to which the formulation will be exposed.Also a coating may be used which is not resistant to low pH values butwhich delays the release of the enzyme at low pH values. It is alsopossible to prepare the agent according to the present invention ascoated (see above) pellets, granules or micro-/mini-tablets which can befilled into coated or non-coated capsules or which can be pressed intocoated or noncoated tablets. Suitable coatings are, for example,cellulose acetate phthalate, cellulose derivates, shellac,polyvinylpyrrolidone derivates, acrylic acid, polyacrylic acid derivatesand polymethyl methacrylate (PMMA), such as e.g. Eudragit® (from RöhmGmbH, Darmstadt, Germany), in particular Eudragit® L30D-55. The coatingEudragit® L30D-55 is dissolved, for example, at a pH value of 5.5 andhigher. If it is desired to release the enzyme already at a lower pHvalue, this may be achieved e.g. by the addition of sodium hydroxidesolution to the coating agent Eudragit® L30D-55, because in this casecarboxyl groups of the methacrylate would be neutralised. Therefore,this coating will be dissolved, for example, already at a pH value of4.0 provided that 5% of the carboxyl groups are neutralised. Theaddition of about 100 g of 4% sodium hydroxide solution to 1 kg ofEudragit® L30D-55 would result in a neutralisation of about 6% of thecarboxyl groups. Further details about formulation methods andadministration methods can be found in the 21st edition of “Remington:The Science & Practice of Pharmacy”, published 2005 by Lippincott,Williams & Wilkins, Baltimore, USA, in the Encyclopedia ofPharmaceutical Technology (Editor James Swarbrick) and in Prof. Bauer“Lehrbuch der Pharmazeutischen Technologie”, 18th edition, published2006 by Wissenschaftliche Verlagsgesellschaft (ISBN 3804-72222-9). Thecontents of these documents are incorporated herein by reference.

Other suitable acceptable carriers or adjuvants for use in the presentinvention include, but are not restricted to water, mineral oil,ethylene glycol, propylene glycol, lanolin, glyceryl stearate, sorbitanstearate, isopropyl myristate, isopropyl palmitate, acetone, glycerine,phosphatidylcholine, sodium cholate or ethanol.

The compositions for use in the present invention may also comprise atleast one co-emulsifying agent which includes but is not limited tooxyethylenated sorbitan monostearate, fatty alcohols, such as stearylalcohol or cetyl alcohol, or esters of fatty acids and polyols, such asglyceryl stearate.

The agents according to the present invention may be provided in astabilized form. Generally, stabilization methods and procedures whichmay be used according to the present invention include any and allmethods for the stabilization of chemical or biological material whichare known in the art, comprising e.g. the addition of chemical agents,methods which are based on temperature modulation, methods which arebased on irradiation or combinations thereof. Chemical agents that maybe used according to the present invention include, among others,preservatives, acids, bases, salts, antioxidants, viscosity enhancers,emulsifying agents, gelatinizers, and mixtures thereof.

Usually, the industrial production of enzymes is performed in atechnical fermentation way using suitable microorganisms (bacteria,moulds, fungi). Usually the strains are recovered from naturalecosystems according to a special screening protocol, isolated as purecultures as well as improved in their properties with respect to theenzyme spectrum and biosynthesis performance (volume/time yield). Enzymeproduction may also be carried out by methods developed in the future.

5-D-fructose dehydrogenase is commercially available (e.g. Sigma-Aldrichor Toyobo Enzymes, Japan) and is usually prepared in a microbiologicalway with the help of the microorganism Gluconobacter industrius. Glucoseisomerase is also commercially available (e.g. Sigma-Aldrich orNovozymes A/S, Denmark) and usually prepared in a microbiological waywith the help of the microorganism Streptomyces murinus. However, theinvention is not limited to the enzymes that are commercially availableat the moment, but generally relates to enzymes that can catalyze theconversion of fructose—specifically or non-specifically—to5-keto-D-fructose, and of glucose—specifically or non-specifically—tofructose. A person skilled in the art can prepare suitable furtherenzymes by conventional methods, for example by mutagenesis of the geneencoding 5-D-fructose dehydrogenase which is present in Gluconobacterindustrius or by mutagenesis of the gene encoding glucose isomerase instreptomyces murinus. The enzymes may also be prepared with the help ofother microorganisms, such as fungi, in sufficient amounts and therequired purities, also by the use of the genetic engineering methodswhich are presently known or may be developed in the future. Forexample, if it is desired to produce the enzymes with othermicroorganisms, then the genetic information of a microorganism whichhas been found initially by extensive screening and which has beenproven to be a suitable source of the enzyme with the desired propertiescan be transferred to a microorganism which is normally used for theproduction of enzymes. Also the modification of the enzyme(s) and theproduction of the enzyme(s) by means of methods which are presentlyknown or may be developed in the future in the area of industrial enzymedevelopment and enzyme production, such as genetic engineering, ispossible. The use and the manner of performing all these methods fordeveloping and producing the enzyme(s) with the desired purities andactivities and with the desired properties, in particular with respectto the stability of the enzyme(s) at various pH values, regarding theoptimum of the pH value, the stability at various temperatures andtemperature optimum, are well known to a person skilled in the art. Theexplanations in chapter 2 (page 82 to page 130) of the textbook“Lebensmittel-Biotechnologie and Ernährung” of Heinz Ruttloff, JürgenProll and Andreas Leuchtenberger, published by Springer Verlag 1997(ISBN 3-540-61135-5) describe these methods in detail. These methods arealso described in “Advances in Fungal Biotechnology for Industry,Agriculture, and Medicine” by Jan S. Tkacz, Lene Langeand (published in2004, ISBN 0-306-47866-8), in “Enzymes in Industry: Production andApplications” by Wolfgang Aehle (Editor), published in 2004, ISBN3527295925 and in “Microbial Enzymes and Biotransformations” byJose-Luis Barredo (Humana Press 2005, ISBN 1588292533). These documentsare herewith incorporated into the patent application by reference. Allthis also applies to the enzymes mentioned below that can optionally beadded to the agent according to the present invention.

The activity of 5-D-fructose dehydrogenase is defined in units (assayavailable e.g. from Sigma-Aldrich), whereby one unit is the amount of5-D-fructose dehydrogenase that converts one micromole of D-fructose to5-keto-D-fructose per minute at pH 4.5 and 37° C. Generally, theactivity of 5-D-fructose dehydrogenase per dose unit should be between10 and 5 million units, preferably between 25 and 2.5 million units andparticularly preferably between 50 and 1 million units.

In the case of the combination agent according to the present inventionwhich also contains glucose isomerase, the composition should containglucose isomerase in an amount of 0.01 to 100,000 GIU, preferably of0.05 to 10,000 GIU and particularly preferably of 0.1 to 1,000 GIU perdose unit. One unit of this enzyme is defined as a glucose isomeraseunit (GIU). One GIU converts 1 g of glucose into fructose at a pH valueof 6.0 and at a temperature of 37° C. from a solution of initially 10%(percent by weight, i.e. 10 g of glucose +90 g of water) in 5 minutes.

The wide range of the above mentioned dosages may be explained by thefact that the agent according to the present invention can be appliedfor many uses, such as the different categories of adiposity and itsaccompanying and resulting diseases, or simply the effort of persons ofnormal weight to limit the intake of calories for preventing weightgain. Furthermore, the different dosages also result from the fact thatstrongly varying amounts of fructose and glucose are administered to thebody, depending on the respective food.

The agent according to the present invention may comprise one or moreadditional enzymes, such as invertase (syn. beta-fructofuranosidase orbeta-fructosidase), lactase (syn. beta-galactosidase), maltase (syn.alpha-glucosidase), alpha-amylase, beta-amylase, glucoamylase,pullulanase, isoamylase, amyloglucosidase, cyclomaltodextringlucantransferase (CGTase). These enzymes have the property of releasingfructose and/or glucose from fructose and/or glucose containingsubstances and foodstuffs—alone or in combination with one or more ofthese enzymes—, whereby the enzymes pullulanase and isoamylase alsoincrease the efficiency of glucoamylase and beta-amylase. All theseenzymes are commercially available (e.g. BioCat Inc., Troy, USA orNovozymes A/S, Denmark or Amano Enzymes Inc., Japan or Sigma-Aldrich)and, up to now, have never been used in combination with 5-D-fructosedehydrogenase—alone or in combination with glucose isomerase—in themedical/pharmaceutical field, in particular not in the case ofadiposity. Thus this application discloses the first medical indicationfor 5-D-fructose dehydrogenase—alone or in combination with glucoseisomerase—in combination with any or all of these enzymes. Examples foragents according to the present invention include: 5-D-fructosedehydrogenase in combination with invertase, or 5-D-fructosedehydrogenase in combination with glucose isomerase and invertase, or5-D-fructose dehydrogenase in combination with glucose isomerase andlactase and invertase, still further 5-D-fructose dehydrogenase incombination with glucose isomerase, Invertase, alpha amylase, betaamylase, glucoamylase, maltase, iso-amylase and pullulanase (combinationof 9 enzymes), or 5-D-fructose dehydrogenase in combination with glucoseisomerase, alpha amylase, beta amylase, glucoamylase, maltase,iso-amylase and pullulanase as well as invertase and lactase(combination of 10 enzymes).

For example, said invertase can release glucose and fructose from e.g.sucrose and said lactase can release glucose from lactose. Beta-amylasebreaks down e.g. 1,4-alpha bonds in starch, starting at the non-reducingend of the polysaccharide chain with cleaving of maltose, and glucose isreleased by the action of maltase on maltose. By the addition of one ormore of these enzymes to the agent according to the present invention,the endogenic release of fructose and/or glucose from fructose- and/orglucose-containing substances or foodstuffs, in particular from sucroseand starch, may also be promoted and accelerated, so that the conversionof fructose into 5-keto-D-fructose which is catalyzed by 5-D-fructosedehydrogenase and/or the conversion of glucose to fructose, which iseffected by glucose isomerase, may occur earlier. Therefore, theaddition of one or more of these enzymes to the agent according to thepresent invention may have the benefit of reducing the required amountof 5-D-fructose dehydrogenase and glucose isomerase.

The activity of invertase is measured in Sumner units (SU, assayavailable e.g. from BioCat Inc., Troy, Va., USA). An SU is defined asthe amount of the enzyme which converts 1 mg of sucrose into glucose andfructose under standard test conditions within 5 minutes at 20° C. and apH value of 4.5. If the agent according to the present invention alsocontains invertase, the activity of the invertase per dose unit shouldbe between 50 and 250,000 SU, preferably between 100 and 150.000 SU andparticularly preferably between 150 and 100,000 SU per dose unit.

The activity of lactase is given in Food Chemical Codex (FCC) units(assay is published in the Food Chemical Codex, fifth edition, and alsoavailable e.g. from Bio Cat Inc. Troy, Va. or Amano Enzymes, Japan orfrom Sigma Aldrich). If the agent according to the present inventionalso contains lactase, the activity of the lactase per dose unit shouldbe between 50 and 200,000 FCC units, preferably between 100 and 100,000FFC units and particularly preferably between 150 and 50,000 FCC units.

The activity of maltase is defined in units, wherein one unit is theamount of maltase which will convert maltose to D-glucose at a rate ofone milligram per minute at 37° C. and a pH of 4.0 in a 10% maltosesolution by weight.

Where the agent according to the present invention also containsmaltase, the activity per dose unit should be between 100 and 100,000units, preferably between 200 and 50,000 units and particularlypreferably between 500 and 20,000 units.

Also for the other enzymes mentioned, the standard test conditions andthe way in which the enzyme activities are to be determined are knownand can be read up by specialists in the field.

Insofar as one or more of the optional enzymes are added to the agentaccording to the present invention, they—as is the case for the5-D-fructose dehydrogenase and the glucose isomerase—should be used insufficient amounts so that they can develop a sufficient enzyme activityfor the intended purpose, e.g. sufficient invertase, so that an amountof sucrose usually ingested with a normal meal (e.g. 15 g) can becleaved, and/or lactase, so that an amount of lactose usually ingestedwith a normal meal (e.g. 10 g) can be cleaved.

The enzymes used can be for example in solid form, e.g. as crystallineor amorphous granules or powders, as a paste or as a liquid, as well asin other forms. In some embodiments, the enzyme is a free enzyme. Inother embodiments, the enzyme may e.g. be immobilized on substrate,which can be powderized if necessary before the enzyme is used inaccordance with the invention.

If the agent according to the present invention is added to a foodstuffbefore eating or during production, the activity of 5-D-fructosedehydrogenase should be between 10 and 250,000 units, preferably between25 and 150,000 units and particularly preferably between 50 and 100,000units per gram fructose in the foodstuff. If the agent also containsglucose isomerase and the agent according to the present invention isadded to a foodstuff before eating or during production, the activity ofglucose isomerase should be between 0.01 and 20,000 units, preferablybetween 0.05 and 10,000 units and particularly preferably between 0.1and 1000 units per gram glucose in the foodstuff.

If the agent according to the present invention is added to a foodstuffbefore eating or during production and if the agent contains5-D-fructose dehydrogenase and glucose isomerase, the activity of5-D-fructose dehydrogenase should be between 10 and 250,000 units,preferably between 25 and 150,000 units and particularly preferablybetween 50 and 100,000 units per gram of fructose and glucose combinedcontained in the foodstuff.

It may be advantageous to add an electron acceptor to the agentaccording to the present invention at e.g. a ratio (acceptor:substrate)of 1:1 to 1:1,000, preferably at a ratio of 1:2 to 1:200, particularlypreferably at a ratio of 1:10 to 1:50. Examples of suitable acceptorswhich may be used include NAD⁺, NADP⁺, FAD⁺, vitamins, such as vitaminC, vitamin E or vitamin A, ferricyanide, ketones, aldehydes,2,6-dichlorophenolindophenol, phenazine methosulfate, nitrobluetetrazolium (including mixtures thereof), but are not limited thereto.

The physiologically present electrolytes should be sufficient for thefunction of the glucose isomerase. But it may also be advantageous toadd electrolytes to the agent according to the present invention, e.g.in an amount of 0.0001% to 0.1% of the substrate (glucose). Examples ofthe electrolytes include, but are not limited to, MgSO₄, Na₂CO₃, NaHCO₃,NaOH, Na₂SO₄, MgCO₃, H₂SO₄, NaS₂O₃, NaS₂O₅ (including mixtures thereof).

It may also be advantageous to add metal ions, in particular cations,such as Mn2+, Mg2+, Ca2+, Zn2+, Fe2+, Co2+ or Cu2+, including mixturesthereof, to the agent according to the present invention, namelypreferably in a molar ratio of 10-6 to 10-2. For the above mentioned(xylose) glucose isomerase which is described by Yamanaka, in particularMn2+ is a suitable cation.

Capsule sizes mentioned below refer to the size definitions used byCapsugel Belgium BVBA, Bornem, Belgium. The size of the capsules shouldbe chosen according to the specific formulation of the agent.

A composition according to the present invention for the production ofcapsules (for example of size 00) may consist of 370 mg of 5-D-fructosedehydrogenase with an activity of 90 units/mg and 100 mg of dicalciumphosphate per capsule.

Another example for a dosage form according to the present inventionconsists of capsules (size 1) that contain 110 mg of 5-D-fructosedehydrogenase with an activity of 500 units/mg as well as 50 mg ofinvertase with an activity of 200 SU units/mg as well as 50 mg ofmaltase with an activity of 200 units/mg and 90 mg of maltodextrin.

Another example for the dosage form according to the present inventionconsists of capsules of size 00 which contain 200 mg of 5-D-fructosedehydrogenase with an activity of 500 units/mg as well as 100 mg ofinvertase with an activity of 200 SU units/mg as well as 150 mg ofdicalcium phosphate.

In the case of a combination agent in which 5-D-fructose dehydrogenaseand glucose isomerase are used, a capsule of size 0 may contain 250 mgof 5-D-fructose dehydrogenase with an activity of 90 units/mg and 20 mgof glucose isomerase with an activity of 1 GIU/mg and 50 mg dicalciumphosphate.

A further example of a combination preparation with 5-D-fructosedehydrogenase and glucose isomerase for the production of capsules ofsize 00 may contain 370 mg of 5-D-fructose dehydrogenase with anactivity of 90 units/mg, 30 mg of glucose isomerase with an activity of1 GIU/mg and 70 mg of dicalcium phosphate.

A further example of a combination preparation with 5-D-fructosedehydrogenase and glucose isomerase for the production of capsules ofsize 00 may contain 110 mg of 5-D-fructose dehydrogenase with anactivity of 500 units/g, 50 mg of glucose isomerase with an activity of1 GIU/mg, 100 mg of invertase with an activity of 200 SU units/mg, 90 mgof lactase with an activity of 100 FCC units/mg and 120 mg of dicalciumphosphate.

A further example of a combination preparation with 5-D-fructosedehydrogenase and glucose isomerase for the production of capsules (e.g.of size 3) may consist of 55 mg of 5-D-fructose dehydrogenase with anactivity of 1000 units/mg, 50 mg of glucose isomerase with an activity 1GIU/mg and 55 mg of dicalcium phosphate per capsule.

A further example for a dosage form according to the present inventionconsists of capsules (size 00) that contain 165 mg of 5-D-Fructosedehydrogenase with an activity of 1000 units/mg, 150 mg of glucoseisomerase with an activity of 1 GIU/mg and 155 mg of dicalcium phosphateper capsule.

The invention may for example contain between 10 and 5 million units of5-D-fructose dehydrogenase per dose unit. In addition, suitableadditives in the required amount may be used. The invention may, inaddition, contain glucose isomerase, for example between 0.01 and100,000 GIU (=glucose isomerase units) per dose unit.

The invention may be provided for medical purposes and non-medicalpurposes, e.g. as a pharmaceutical composition, medical device,foodstuff or special foodstuff.

With the agents according to the present invention the content ofcalories of carbohydrate containing food that can be utilized by thebody can be reduced to such a degree that a marked weight loss can beachieved or a weight gain can be prevented despite the intake ofcarbohydrate rich food. The agents according to the present inventionare thus suitable for use in methods for the therapeutic treatment ofthe human or animal body in which a limitation or reduction of theintake of food calories originating from carbohydrates is intended, e.g.in the case of the therapy of diseases of the cardiovascular system(e.g. hypertension, coronary heart disease, venous insufficiency, heartfailure, left-ventricular hypertrophy, arteriosclerosis), of metabolicand hormonal function (e.g. diabetes mellitus type II, dyslipidemias,hyperuricemia, hyperlipoproteinemia), of the respiratory system (e.g.sleep apnea, Pickwickian syndrome), of the hepatobiliary system (e.g.fatty liver, cholecystolithiasis), of the locomotor system (e.g.gonarthrosis, heel spur, arthrosis of the ankle joint), of the skin(e.g. intertrigo, hirsutism, striae), in the therapy of neoplasias (e.g.increased risk of endometrial, breast, cervical, gall bladder cancer,etc.), in the therapy of disorders of sexual function (e.g. reducedfertility, complications during birth), in the therapy of psychosocialproblems (e.g. reduced self-confidence, social isolation,discrimination, problems with one's partner or at work) and otherproblems, such as reduced mobility and staying power, increased riskduring operations and more difficult examination conditions. All thesediseases and health problems are often associated with adiposity.

The agents according to the present invention are in particular suitablefor use in the case of adiposity and for the therapeutic treatment ofadiposity.

In the description and following claims, the term “fructoseequivalent-containing” refers to all substances and foodstuffs thatcontain fructose (a) as fructose per se, (b) in a form from whichfructose can be released in the digestive tract (e.g. by cleavage asfructose from a saccharide chain containing at least two saccharidemonomers), (c) in a form that can be converted to fructose, e.g. asglucose per se, or (d) in a form that can be released in the digestivetract and converted to fructose, e.g. as a saccharide chain containingat least two saccharide monomers, at least one of which can be cleavedfrom the saccharide chain as glucose.

In the description and following claims, the term “total fructose”refers to the total content of fructose in a foodstuff (a) as fructoseper se, (b) in a form from which fructose can be released in thedigestive tract (e.g. by cleavage as fructose from a saccharide chaincontaining at least two saccharide monomers), (c) in a form that can beconverted to fructose, e.g. as glucose per se, or (d) in a form that canbe released in the digestive tract and converted to fructose, e.g. asaccharide chain containing at least two saccharide monomers, at leastone of which can be cleaved from the saccharide chain as glucose.

In the description and following claims, the term “effective fructosecontent” of an item refers to the effective amount of total fructose inthat item, taking into account the prior action of fructose convertingenzymes that have been added to the item or the future action offructose converting enzymes that have been added to the item. Thus, forexample, a foodstuff having a given fructose content and havingmicroencapsulated 5-D-fructose dehydrogenase incorporated therein willhave a lower effective fructose content than a foodstuff which lacks themicroencapsulated 5-D-fructose dehydrogenase but is otherwise identical,since release of the 5-D-fructose dehydrogenase after ingestion willresult in at least a portion of the fructose in the foodstuff beingconverted to 5-keto D-fructose.

Thus, there is provided, in accordance with embodiments of theinvention, a 5-D-fructose dehydrogenase, optionally in combination withone or more enzyme(s) selected from glucose isomerase, invertase,lactase, maltase, alpha-amylase, beta-amylase, glucoamylase,pullulanase, isoamylase, amyloglucosidase and cyclomaltodextringlucanotransferase, for use in medicine.

There is also provided, in accordance with embodiments of the invention,a pharmaceutical composition comprising 5-D-fructose dehydrogenaseoptionally in combination with one or more enzyme(s) selected fromglucose isomerase, invertase, lactase, maltase, alpha-amylase,beta-amylase, glucoamylase, pullulanase, isoamylase, amyloglucosidaseand cyclomaltodextrin glucanotransferase.

In some embodiments, at least one of the enzymes, or the pharmaceuticalcomposition, is protected by a coating to be stable at pH values of lessthan 4, preferably less than 3. In some embodiments, the composition ofmatter as described above is in a form for oral administration. In someembodiments, the composition of matter as described above is in a formsuited to be added to food at the production stage of the same and/orbefore eating.

There is also provided, in accordance with embodiments of the invention,a medical device comprising 5-D-fructose dehydrogenase optionally incombination with one or more enzyme(s) selected from glucose isomerase,invertase, lactase, maltase, alpha-amylase, beta-amylase, glucoamylase,pullulanase, isoamylase, amyloglucosidase and cyclomaltodextringlucanotransferase. In some embodiments, the medical device includes oneor more enzyme(s) protected by a coating to be stable at pH values ofless than 4, preferably less than 3. In some embodiments, the medicaldevice is in a form for oral administration. In some embodiments, themedical device is in a form suited to be added to food at the productionstage of the same and/or before eating.

There is also provided, in accordance with embodiments of the invention,a foodstuff comprising 5-D-fructose dehydrogenase in combination withinvertase and/or maltase and/or lactase.

There is also provided, in accordance with embodiments of the invention,a special foodstuff comprising 5-D-fructose dehydrogenase optionally incombination with one or more enzyme(s) selected from glucose isomerase,invertase, lactase, maltase, alpha-amylase, betaamylase, glucoamylase,pullulanase, isoamylase, amyloglucosidase and cyclomaltodextringlucanotransferase. In some embodiments, the special foodstuff includesone or more enzyme(s) protected by a coating to be stable at pH valuesof less than 4, preferably less than 3. In some embodiments, the specialfoodstuff is in a form for oral administration. In some embodiments, thespecial foodstuff according is in a form suited to be added to food atthe production stage of the same and/or before eating.

In some embodiments, the pharmaceutical composition, medical device orthe special foodstuff as described above comprises 5-D-fructosedehydrogenase in combination with glucose isomerase.

There is also provided, in accordance with embodiments of the invention,the use of 5-D-fructose dehydrogenase, optionally in combination withone or more enzyme(s) selected from glucose isomerase, invertase,lactase, maltase, alpha-amylase, beta-amylase, glucoamylase,pullulanase, isoamylase, amyloglucosidase and cyclomaltodextringlucanotransferase, for the production of an agent, preferably apharmaceutical composition, for the curative or prophylactic treatmentof adiposity. In some embodiments, the adiposity is accompanied bydisturbances of the cardiovascular system and/or of the metabolic andhormone based function and/or of the respiratory system and/or of thehepatobiliary system and/or disturbances of the locomotor system and/ordiseases of the skin and/or neoplasias and/or disorders of the sexualfunction and/or psychosocial problems and/or reduced mobility andstaying power and/or an increased operation risk and/or more difficultexamination conditions. In some embodiments, the disturbances of thecardiovascular system are hypertension, coronary heart disease, venousinsufficiency, heart failure, left-ventricular hypertrophy and/orarteriosclerosis. In some embodiments, the disturbances of the metabolicor hormonal function are diabetes mellitus type II, dyslipidemias,hyperuricemia and/or hyperlipoproteinemia. In some embodiments, thedisturbances of the respiratory system are sleep apnea and/or thePickwickan syndrome. In some embodiments, the disturbances of thehepatobiliary system are fatty liver and/or cholecystolithiasis. In someembodiments, the disturbances of the locomotor system are gonarthrosis,heel spur and/or arthrosis of the ankle joint. In some embodiments, thedisturbances of the skin are intertrigo, hirsutism and/or striae. Insome embodiments, the neoplasias are endometrial, mama, cervix and/orgall bladder carcinoma. In some embodiments, the disorders of the sexualfunction are reduced fertility and/or an increased risk of complicationsin the case of birth. In some embodiments, the psychosocial problems arereduced self-confidence, social isolation, discrimination and/or,problems with the partner and/or at the job.

There is also provided, in accordance with embodiments of the invention,the use of 5-D-fructose dehydrogenase, optionally in combination withone or more enzyme(s) selected from glucose isomerase, invertase,lactase, maltase, alpha-amylase, beta-amylase, glucoamylase,pullulanase, isoamylase, amyloglucosidase and cyclomaltodextringlucanotransferase for production of an agent, preferably apharmaceutical composition, for lowering the bioavailability of fructoseand/or glucose in the human or animal body.

There is also provided, in accordance with embodiments of the invention,the use of 5-D-fructose dehydrogenase, optionally in combination withone or more enzyme(s) selected from glucose isomerase, invertase,lactase, maltase, alpha-amylase, beta-amylase, glucoamylase,pullulanase, isoamylase, amyloglucosidase and cyclomaltodextringlucanotransferase for production of an agent, preferably apharmaceutical composition, for lowering the content of fructose and/orglucose in a foodstuff.

There is also provided, in accordance with embodiments of the invention,the use of 5-D-fructose dehydrogenase, optionally in combination withone or more enzyme(s) selected from glucose isomerase, invertase,lactase, maltase, alpha-amylase, beta-amylase, glucoamylase,pullulanase, isoamylase, amyloglucosidase and cyclomaltodextringlucanotransferase for production of an agent, preferably apharmaceutical composition, for lowering the usable calorie content offood.

In some embodiments, in the use described above, the agent, preferablythe pharmaceutical composition, is in a form for oral administration.

In some embodiments, in the use described above, at least two enzymesare present. In some embodiments, the two enzymes are 5-D-fructosedehydrogenase and glucose isomerase.

In some embodiments, in the use described above, the agent is selectedfrom a pharmaceutical composition, a medical device, a foodstuff or aspecial foodstuff.

In some embodiments, in the use described above, the enzymes areprotected by a coating to be stable at pH values of less than 4,preferably less than 3.

In some embodiments, in the use described above, the agent is in a formfor oral use.

In some embodiments, in the use described above, the product is suitedto be added to food at the production stage of the same and/or beforeeating.

In some embodiments, in the use described above, the product is in aform for use in immobilised form.

There is also provided, in accordance with embodiments of the invention,a process for the treatment of a foodstuff, comprising the steps ofcontacting the foodstuff with a 5-D-fructose dehydrogenase optionally incombination with one or more enzyme(s) selected from glucose isomerase,invertase, lactase, maltase, alpha-amylase, beta-amylase, glucoamylase,pullulanase, isoamylase, amyloglucosidase and cyclomaltodextringlucanotransferase, and initiating the reduction of the fructose contentand optionally the reduction of the glucose content of the foodstuff. Insome embodiments, as a further step prior to the initiation, ingestionof the foodstuff takes place.

There is also provided, in accordance with embodiments of the invention,a mammalian-ingestible composition of matter which comprises an enzymethat converts D-fructose into a form that is biologically inactive in achosen mammalian body or a mixture of such enzymes.

There is also provided, in accordance with embodiments of the invention,a mammalian-ingestible composition of matter which comprises an enzymethat converts D-fructose into a form that cannot be digested in a chosenmammalian digestive tract.

There is also provided, in accordance with embodiments of the invention,a mammalian-ingestible composition of matter which comprises an enzymethat converts D-fructose into a form that is not metabolizable in achosen mammalian body.

In some embodiments, the chosen mammalian body is a human body. In someembodiments, the chosen mammalian body is a non-human body. In someembodiments, the form into which D-fructose is converted is5-keto-D-fructose. In some embodiments, the enzyme is 5-D-fructosedehydrogenase. In some embodiments, the composition of matter is a humandietary supplement or a pharmaceutical composition. In some embodiments,the composition of matter is an animal dietary supplement or aveterinary composition. In some embodiments, the composition is aspecial foodstuff. In some embodiments, the composition of matterfurther comprises at least one pharmaceutically or dietarily acceptablecarrier or excipient. In some embodiments, the composition of matterfurther comprises at least one veterinarily acceptable carrier orexcipient. In some embodiments, the composition of matter contains theenzyme in microencapsulated form. In some embodiments, the compositionof matter is in the form of a capsule or tablet. In some embodiments,the composition of matter is in the form of granules or pellets. In someembodiments, the composition of matter is in the form of a solution. Insome embodiments, the composition of matter is in the form of a liquid.In some embodiments, the composition of matter is in the form of a gelor suspension. In some embodiments, the composition of matter is in theform of a gelcap. In some embodiments, the composition of matter is inthe form of a powder.

There is also provided, in accordance with embodiments of the invention,a composition of matter which is microencapsulated enzyme 5-D-fructosedehydrogenase.

In some embodiments, the composition of matter as described above isadapted to be mixed with a food.

There is also provided, in accordance with embodiments of the invention,a composition of matter comprising the enzyme 5-D-fructose dehydrogenaseadmixed with a mammalian-ingestible substance. In some embodiments, themammalian-ingestible substance is a human-ingestible substance. In someembodiments, the mammalian-ingestible substance is an animal-ingestiblesubstance. In some embodiments, the mammalian-ingestible substance is apharmaceutically or dietarily acceptable carrier or excipient. In someembodiments, the mammalian-ingestible substance is a veterinarilyacceptable carrier or excipient. In some embodiments, the 5-D-fructosedehydrogenase is microencapsulated.

In some embodiments, in the composition of matter as described above,the enzyme constitutes between 5 and 99.9% by weight of the compositionof matter. In some embodiments, the enzyme constitutes between 10 and80% by weight of the composition of matter. In some embodiments, theenzyme constitutes between 25 and 60% by weight of the composition ofmatter.

In some embodiments, the composition of matter as described above is inunit dosage form and the unit dosage contains between 10 and 5 millionunits of 5-D-fructose dehydrogenase activity. In some embodiments, theunit dosage contains between 25 and 2 5 million units of 5-D-fructosedehydrogenase activity. In some embodiments, the unit dosage containsbetween 50 and 1 million units of 5-D-fructose dehydrogenase activity.

In some embodiments, the composition of matter described above comprisesa coating which dissolves in an aqueous medium at a pH of between 3.0and 8.0. In some embodiments, the coating does not dissolve in anaqueous medium at a pH of below 3.0. In some embodiments, the coatingdoes not dissolve in an aqueous medium at a pH below 4.0. In someembodiments, the coating does not dissolve in an aqueous medium at a pHabove 6.5. In some embodiments, the coating does not dissolve in anaqueous medium at a pH above 6.0.

In some embodiments, the composition of matter as described above is aslow-release or extended-release formulation. In some embodiments, theslow-release or extended-release formulation comprises a slow-release orextended-release coating.

In some embodiments, the composition of matter as described abovefurther comprises a second enzyme. In some embodiments, the secondenzyme is capable of cleaving fructose or glucose from a sugar thatcontains at least two saccharide monomers. In some embodiments, theenzyme is invertase or maltase. In some embodiments, the second enzymeis invertase, the composition of matter is in unit dosage form, and eachunit dosage contains between 50 and 250,000 Sumner units of invertaseactivity. In some embodiments, each unit dosage contains between 100 and150,000 Sumner units of invertase activity. In some embodiments, eachunit dosage contains between 150 and 100,000 Sumner units of invertaseactivity. In some embodiments, the second enzyme is maltase, thecomposition of matter is in unit dosage form, and each unit dosagecontains between 100 and 100,000 units of maltase activity. In someembodiments, each unit dosage contains between 200 and 50,000 units ofmaltase activity. In some embodiments, each unit dosage contains between500 and 20,000 units of maltase activity.

In some embodiments, the composition of matter as described abovecomprises both invertase and maltase.

In some embodiments, the composition of matter as described abovefurther comprises an additional enzyme that converts D-fructose into amolecule that is absorbed more quickly than D-fructose from theintestine into the bloodstream. In some embodiments, the molecule isD-glucose.

In some embodiments, the composition of matter as described abovefurther comprises an additional enzyme that converts D-glucose toD-fructose. In some embodiments, the additional enzyme is a glucoseisomerase. In some embodiments, the composition of matter is in unitdosage form and each dosage unit contains 0.01 to 100,000 units ofglucose isomerase activity per dose unit. In some embodiments, eachdosage unit contains 0.05 to 10,000 units of glucose isomerase activityper dose unit. In some embodiments, each dosage unit contains 0.1 to1,000 units of glucose isomerase activity per dose unit. In someembodiments, the glucose isomerase is a xylose isomerase.

In some embodiments, the composition of matter as described abovefurther comprises one or more members of the group consisting oflactase, alpha-amylase, beta-amylase, glucoamylase, pullulanase,isoamylase, amyloglucosidase and cyclomaltodextrin glucantransferase(CGTase). In some embodiments, the member is lactase, the composition ofmatter is in unit dosage form and each dosage unit contains 50 to200,000 FCC units of lactase activity per dose unit. In someembodiments, each dosage unit contains 100 to 100,000 FCC units oflactase activity per dose unit. In some embodiments, each dosage unitcontains 150 to 50,000 units of lactase activity per dose unit.

In some embodiments, the composition of matter further comprises atleast one of an electrolyte and a metal ion. In some embodiments, theelectrolyte is selected from the group consisting of MgSO₄, Na₂CO₃,NaHCO₃, NaOH, Na₂SO₄, MgCO₃, H₂SO₄, NaS₂O₃, NaS₂O₅ and mixtures thereof.In some embodiments, the metal ion is selected from the group consistingof Mn²⁺, Mg²⁺, Ca²⁺, Zn²⁺, Fe²⁺, Co²⁺, Cu²⁺ and mixtures thereof.

In accordance with some embodiments, the composition of matter is afoodstuff, and the enzyme is in active form. In some embodiments, theamount or concentration of the enzyme in the foodstuff is greater thanthe naturally occurring concentration or amount of the enzyme in thefoodstuff. In some embodiments, the foodstuff is a fructoseequivalent-containing foodstuff. In some embodiments, the foodstuff is afructose-containing foodstuff. In some embodiments, the foodstuff is aglucose-containing foodstuff. In some embodiments, the enzyme is5-D-fructose dehydrogenase which is present in an amount of 10 to250,000 units of activity per gram of fructose in the foodstuff. In someembodiments, the 5-D-fructose dehydrogenase is present in an amount of25 and 150,000 units of activity per gram of fructose in the foodstuff.In some embodiments, the 5-D-fructose dehydrogenase is present in anamount of 50 to 100,000 units of activity per gram of fructose in thefoodstuff. In some embodiments, the foodstuff is a glucose-containingfoodstuff and contains glucose isomerase in an amount of 0.01 to 20,000units of activity per gram of glucose in the foodstuff. In someembodiments, the glucose isomerase is present in an amount of 0.05 to10,000 units of activity per gram of glucose in the foodstuff. In someembodiments, the glucose isomerase is present in an amount of 0.1 to1,000 units of activity per gram of glucose in the foodstuff. In someembodiments, the enzyme is 5-D-fructose dehydrogenase which is presentin an amount of 10 to 250,000 units of activity per gram of totalfructose in the foodstuff. In some embodiments, the 5-D-fructosedehydrogenase is present in an amount of 25 and 150,000 units ofactivity per gram of total fructose in the foodstuff. In someembodiments, the 5-D-fructose dehydrogenase is present in an amount of50 to 100,000 units of activity per gram of total fructose in thefoodstuff.

In some embodiments, the foodstuff is a foodstuff which has been baked.In some embodiments, the foodstuff is a foodstuff which has been cooked.In some embodiments, the foodstuff is a liquid, paste or broth. In someembodiments, the enzyme is present in the foodstuff in microencapsulatedform. In some embodiments, the enzyme is 5-D-fructose dehydrogenase. Insome embodiments, the foodstuff further contains a second enzyme inactive form. In some embodiments, the concentration or amount of thesecond enzyme in the foodstuff is greater than the naturally occurringconcentration or amount of the second enzyme in the foodstuff. In someembodiments, the second enzyme is selected from the group consisting ofinvertase, maltase, a glucose isomerase, lactase, alpha-amylase,beta-amylase, glucoamylase, pullulanase, isoamylase, amyloglucosidaseand cyclomaltodextrin glucantransferase (CGTase) or a mixture thereof.In some embodiments, the second enzyme is invertase. In someembodiments, the second enzyme is maltase. In some embodiments, thesecond enzyme is lactase. In some embodiments, the second enzyme isalpha-amylase. In some embodiments, the second enzyme is beta-amylase.In some embodiments, the second enzyme is glucoamylase. In someembodiments, the second enzyme is pullulanase. In some embodiments, thesecond enzyme is isoamylase. In some embodiments, the second enzyme isamyloglucosidase. In some embodiments, the second enzyme is CGTase. Insome embodiments, the second enzyme is a glucose isomerase. In someembodiments, the glucose isomerase is a xylose isomerase. In someembodiments, the composition of matter further comprises at least one ofan electrolyte and a metal ion. In some embodiments, the electrolyte isselected from the group consisting of MgSO₄, Na₂CO₃, NaHCO₃, NaOH,Na₂SO₄, MgCO₃, H₂SO₄, NaS₂O₃, NaS₂O₅ and mixtures thereof. In someembodiments, the metal ion is selected from the group consisting ofMn²⁺, Mg²⁺, Ca²⁺, Zn²⁺, Fe²⁺, Co²⁺, Cu²⁺ and mixtures thereof. In someembodiments, the second enzyme is a mixture of at least two of the groupof invertase, maltase and a glucose isomerase. In some embodiments, thesecond enzyme is a mixture of at least two of the group of invertase,maltase, a glucose isomerase, lactase, amylase, beta-amylase,glucoamylase, pullulanase, isoamylase, amyloglucosidase and CGTase. Insome embodiments, the second enzyme is in microencapsulated form.

In accordance with some embodiments, the composition of matter asdescribed above comprises a mixture of enzymes that convert D-fructoseinto a form that is at least one of (a) biologically inactive in achosen mammalian body, (b) not digestible in the digestive tract of saidmammalian body and (c) not metabolizable in said mammalian body.

In accordance with some embodiments, the composition of matter asdescribed above is a foodstuff which is not a dough.

In accordance with some embodiments, the enzyme that converts D-fructoseinto a form that is at least one of (a) biologically inactive in achosen mammalian body, (b) not digestible in the digestive tract of saidmammalian body and (c) not metabolizable in said mammalian body is notcontained in an inorganic-based sol-gel biocompatible matrix.

In accordance with some embodiments, the composition of matter issubstantially free of substances which are not approved for oral humaningestion. In accordance with some embodiments, the composition ofmatter is substantially free of substances which are not approved fororal non-human mammal ingestion.

In some embodiments, the composition of matter is adapted for oralingestion.

In some embodiments, the composition of matter comprises an electronacceptor. In some embodiments, the electron acceptor is selected fromthe group consisting of Nicotinamide Adenine Dinucleotide+(NAD+),nicotinamide adenine dinucleotide phosphate+(NADP+), flavin adeninedinucleotide+(FAD+), vitamin C, E or A, ferricyanide, ketones,aldehydes, 2,6-di-chloro-phenolindophenol, phenazine methsulfate andmixtures thereof. In some embodiments, the molar ratio of electronacceptor to total fructose is from 1:1 to 1:1000. In some embodiments,the molar ratio of electron acceptor to total fructose is from 1:2 to1:200. In some embodiments, the molar ratio of electron acceptor tototal fructose is from 1:10 to 1:50. In some embodiments, the molarratio of electron acceptor to fructose is from 1:1 to 1:1000. In someembodiments, the molar ratio of electron acceptor to fructose is from1:2 to 1:200. In some embodiments, the molar ratio of electron acceptorto fructose is from 1:10 to 1:50.

In accordance with some embodiments, the composition of matter furthercomprises one or more enzyme stabilizers. In some embodiments, theenzyme stabilizer stabilizes 5-D-fructose dehydrogenase. In someembodiments, the stabilizer stabilizes a glucose isomerase.

There is also provided, in accordance with embodiments of the invention,a method of treating adiposity in a mammalian subject body, comprisingadministering to a mammalian subject an efficacious amount of an enzymeor a mixture of enzymes that converts D-fructose into a form that is atleast one of (a) biologically inactive in the subject body, (b) notdigestible in the subject digestive tract and (c) not metabolizable inthe subject body. There is also provided, in accordance with embodimentsof the invention, a method of reducing the effect of D-fructose on amammalian subject body, comprising administering to a mammalian subjectan efficacious amount of an enzyme or a mixture of enzymes that convertsD-fructose into a form that is at least one of (a) biologically inactivein the subject body, (b) not digestible in the subject digestive tractand (c) not metabolizable in the subject body. There is also provided,in accordance with embodiments of the invention, a method of reducingthe effect of total fructose on a mammalian subject body, comprisingadministering to a mammalian subject an efficacious amount of an enzymeor a mixture of enzymes that converts total fructose into a form that isat least one of (a) biologically inactive in the subject body, (b) notdigestible in the subject digestive tract and (c) not metabolizable inthe subject body. In some embodiments, the mammalian subject is a humansubject. In some embodiments, the mammalian subject is a non-humansubject. In some embodiments, the administering comprises administeringa mammalian-ingestible composition of matter as described above. In someembodiments, the effect of D-fructose or total fructose is adiposity. Insome embodiments, the effect of D-fructose or total fructose is caloricintake via fructose metabolism. In some embodiments, the effect ofD-fructose or total fructose is caloric intake via glucose metabolism.In some embodiments, the effect of D-fructose or total fructose isselected from the group consisting of (a) weight gain (b) accelerationof weight gain (c) the prevention of weight loss and (d) a lessening ofthe rate of weight loss. In some embodiments, the effect of D-fructoseor total fructose is a deleterious effect on the weight or health ofsaid mammal. In some embodiments, the adiposity is treated post-facto.In some embodiments, the adiposity is reduced or preventedprophylactically. In some embodiments, the composition of matter isadministered prior to eating. In some embodiments, the composition ofmatter is administered immediately prior to eating. In some embodiments,the composition of matter is administered concurrently with a meal. Insome embodiments, the composition of matter is administered aftereating. In some embodiments, the composition of matter is administeredimmediately after eating. In some embodiments, the form is into whichD-fructose is converted is 5-keto-D-fructose. In some embodiments, themethod is used as part of a program of therapeutic or prophylactictreatment of at least one of the following, alone or attendant toadiposity: cardiovascular disease, a disease or disorder of metabolicfunction, a disease or disorder of hormonal function, respiratorydisease or disorder, disease or disorder of the hepatobiliary system,disease or disorder of the locomotor system, disease or disorder of theskin, neoplasias associated with adiposity, disorder of sexual function,psychosocial problems, reduced mobility, reduced stamina, and fatigue.In some embodiments, the cardiovascular disease is selected from thegroup consisting of hypertension, coronary heart disease, venousinsufficiency, heart failure, left-ventricular hypertrophy andarteriosclerosis. In some embodiments, the disease or disorder ofmetabolic function is selected from the group consisting of diabetesmellitus type II, dyslipidemias, hyperuricemia and hyperlipoproteinemia.In some embodiments, the respiratory disease or disorder is sleep apneaor Pickwickan syndrome. In some embodiments, the disease or disorder ofthe hepatobiliary system is fatty liver or cholecystolithiasis. In someembodiments, the disease or disorder of the locomotor system is selectedfrom the group consisting of gonarthrosis, heel spur and arthrosis ofthe ankle joint. In some embodiments, the disease or disorder of theskin is selected from the group consisting of intertrigo, hirsutism andstriae. In some embodiments, the neoplasias are selected from the groupconsisting of endometrial, mama, cervix and gall bladder carcinoma. Insome embodiments, the disorder of sexual function is reduced fertilityor an increased risk of complications during birth. In some embodiments,the psychosocial problems are selected from the group consisting ofreduced self-confidence, social isolation, discrimination, problems withthe partner and problems at the job. In some embodiments, the methodfurther comprises administering to the subject a second enzyme selectedfrom the group consisting of invertase, maltase, a glucose isomerase,lactase, alpha-amylase, beta-amylase, glucoamylase, pullulanase,isoamylase, amyloglucosidase and cyclomaltodextrin glucantransferase(CGTase) or a mixture thereof.

There is also provided, in accordance with embodiments of the invention,a kit comprising an enzyme that converts D-fructose into a form that isat least one of (a) biologically inactive in a chosen mammalian body,(b) not digestible in the digestive tract of said mammalian body and (c)not metabolizable in said mammalian body, or a mixture of such enzymes,and instructions explaining how to use said enzyme or mixture thereof toreduce the effects of fructose in said mammalian body. In someembodiments, the instructions explain how to use said enzyme or mixturethereof to reduce the effects of total fructose in said mammalian body.In some embodiments, the mammalian body is the human body. In someembodiments, the mammalian body is a non-human body. In someembodiments, the enzyme or mixture thereof is present as a compositionof matter as described above. In some embodiments, the enzyme or mixtureof enzymes converts D-fructose into 5-keto-D-fructose. In someembodiments, the enzyme is 5-D-fructose dehydrogenase. In someembodiments, the kit further comprises a second enzyme. In someembodiments, the second enzyme is selected from the group consisting ofinvertase, maltase, a glucose isomerase, lactase, alpha-amylase,beta-amylase, glucoamylase, pullulanase, isoamylase, amyloglucosidaseand cyclomaltodextrin glucantransferase (CGTase) or a mixture thereof.In some embodiments, the second enzyme is a glucose isomerase. In someembodiments, the instructions further explain how to use the secondenzyme or mixture thereof in conjunction with the enzyme. In someembodiments, the instructions explain how to use a mixture of the secondenzyme and the enzyme.

1-262. (canceled)
 263. A mammalian ingestible composition which isadapted for oral administration selected from a pharmaceuticalcomposition and a dietary supplement, said composition being in unitdosage form, said composition comprising 5-D-fructose dehydrogenase anda carrier or excipient that is acceptable for use in pharmaceuticalcompositions or foodstuffs, said composition further comprising a secondenzyme selected from the group consisting of a glucose isomerase,maltase, invertase, lactase, alpha-amylase, beta-amylase, glucoamylase,pullulanase, isoamylase, amyloglucosidase, cyclomaltodextringlucanotransferase, and mixtures thereof. 264-266. (canceled)
 267. Thecomposition according to claim 263 which is in a form selected from (a)the group consisting of a tablet, capsule, gel cap, pellet and dragee,(b) powder or (c) liquid form as solution, drops, suspension or gel.268. The composition according to claim 263 wherein said 5-D-fructosedehydrogenase is protected by a coating which is stable at pH below 4.269. The composition of according to claim 268 wherein said coatingprotects the entire dosage unit.
 270. The composition according to claim263 wherein said 5-D-fructose dehydrogenase is microencapsulated. 271.The composition according to claim 263 wherein said 5-D-fructosedehydrogenase constitutes between 5 and 99.9% by weight of thecomposition.
 272. The composition according to claim 263 wherein saidunit dosage contains between 50 and 1 million units of 5-D-fructosedehydrogenase activity.
 273. The composition according to claim 263wherein said composition comprises a coating which dissolves at a pH of5.5 or higher.
 274. The composition according to claim 263 wherein said5-D-fructose dehydrogenase is not contained in an inorganic-basedsol-gel biocompatible matrix.
 275. The composition according to claim263 wherein said second enzyme is a glucose isomerase.
 276. Thecomposition according to claim 275, further comprising a third enzymeselected from the group consisting of invertase, maltase, lactase,alpha-amylase, beta-amylase, glucoamylase, pullulanase, isoamylase,amyloglucosidase, cyclomaltodextrin glucanotransferase and mixturesthereof.
 277. The composition according to claim 263, wherein saidsecond enzyme is selected from the group consisting of lactase,alpha-amylase, beta-amylase, glucoamylase, pullulanase, isoamylase,amyloglucosidase, cyclomaltodextrin glucanotransferase and mixturesthereof.
 278. A foodstuff which comprises 5-D-fructose dehydrogenasewherein at least some of said 5-D-fructose dehydrogenase is present insaid foodstuff in a form in which said 5-D-fructose dehydrogenase willbe released in active form to act on fructose which is present in orreleased from said foodstuff after ingestion of said foodstuff.
 279. Afoodstuff according to claim 278 which further comprises a second enzymeselected from the group consisting of a glucose isomerase, invertase,maltase, lactase, alpha-amylase, beta-amylase, glucoamylase,pullulanase, isoamylase, amyloglucosidase, cyclomaltodextringlucanotransferase and mixtures thereof, wherein at least one of thefollowing is true: (a) at least some of said second enzyme is present insaid foodstuff in active form in which said second enzyme can act on asubstrate which is present in or released from said foodstuff beforeingestion of said foodstuff, and (b) at least some of said second enzymeis present in said foodstuff in a form in which said second enzyme willbe released in active form to act on a substrate which is present in orreleased from said foodstuff after ingestion of said foodstuff. 280-281.(canceled)
 282. A foodstuff according to claim 279, wherein saidfoodstuff is selected from the group consisting of a baked foodstuff, acooked foodstuff, and a liquid foodstuff.
 283. A foodstuff according toclaim 279 wherein said 5-D-fructose dehydrogenase is not contained in aninorganic-based sol-gel biocompatible matrix.
 284. A foodstuff accordingto claim 279 wherein said second enzyme is a glucose isomerase.
 285. Afoodstuff according to claim 284 wherein at least one of said5-D-fructose dehydrogenase and said glucose isomerase ismicroencapsulated.
 286. (canceled)
 287. A method of treating adiposity,reducing the effect of adiposity on a mammalian body, reducing theeffect of D-fructose in a mammalian body, or reducing the effect oftotal fructose in a mammalian body, comprising administering to asubject in need of such treatment or reduction an efficacious amount of5-D-fructose dehydrogenase.
 288. A method according to claim 287,further comprising administering to said subject a second enzymeselected from the group consisting of a glucose isomerase, invertase,maltase, lactase, alpha-amylase, beta-amylase, glucoamylase,pullulanase, isoamylase, amyloglucosidase, cyclomaltodextringlucanotransferase and mixtures thereof. 289-310. (canceled)